Structure of equine type I and type II collagens.
Abstract: Collagen type I was purified from equine skin and flexor tendon, and type II collagen was purified from equine articular cartilage. The proteoglycans in these tissues were extracted, using guanidine HCl; the collagens were solubilized, using pepsin digestion, then were selectively precipitated with NaCl. Gel electrophoresis indicated that the precipitates contained only type I or type II collagen. Amino acid analysis indicated that collagen constituted > 97% of the total protein in the precipitates. Hydroxylation of proline was 42.0 +/- 0.6% (mean +/- SEM) in alpha 1(I) and alpha 2(I), and was 48.1 +/- 1.3% in alpha 1(II) chains. The hydroxylation of lysine was 23.2 +/- 0.7% in alpha 1(I) and 34.1 +/- 0.9% in alpha 2(I) chains from tendon, and 49.6 +/- 4.3% in alpha 1(II) chains from cartilage. The cyanogen bromide (CB)-peptide patterns of chromatographically purified equine alpha 2(I) and alpha 1(II) chains were similar to those published previously for rat, bovine, and human alpha 2 and alpha 1 chains. However, the CB-peptide pattern of the equine alpha 1(I) chain resembled the guinea pig alpha 1(I) chain, which has no methionine between CB7 and CB6. Purified equine alpha 1(I)CB7,6 contained no methionine, methionine sulfoxide, or homoserine lactone. Mass of 42.26 kd was determined by use of mass spectrometry, and N-terminal sequence analysis established that the first 12 amino acids of this CB7,6 were identical to the sequence of human alpha 1(I)CB7.(ABSTRACT TRUNCATED AT 250 WORDS)
Publication Date: 1994-03-01 PubMed ID: 8192271
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The study involves analyzing the structure of type I and type II collagens in horses. The research process involved purification of these collagen types from various tissues and the examination of their characteristics in relation to amino acid components and hydroxylation percentages.
Purification Process
- The researchers purified collagen type I from the skin and flexor tendon of the horse and collagen type II from the equine articular cartilage.
- To extract proteoglycans from the tissues, guanidine HCl was utilized.
- Pepsin digestion served to solubilize the collagens which were then selectively precipitated using sodium chloride (NaCl).
Analysis Results
- When the researchers conducted gel electrophoresis, it was established that the precipitates only contained either type I or type II collagen.
- After the analysis of amino acids, they found that more than 97% of the total protein in the precipitates was constituted by collagen.
- In terms of hydroxylation of proline, the values were 42.0 ± 0.6% for alpha 1(I) and alpha 2(I), and 48.1 ± 1.3% for alpha 1(II) chains.
- For the hydroxylation of lysine, it was 23.2 ± 0.7% in alpha 1(I) and 34.1 ± 0.9% in alpha 2(I) chains from tendon, and 49.6 ± 4.3% in alpha 1(II) chains from cartilage.
Further Comparisons
- The cyanogen bromide (CB)-peptide patterns of equine alpha 2(I) and alpha 1(II) chains after purification were found to be similar to those already published for rat, bovine, and human alpha 2 and alpha 1 chains.
- However, the CB-peptide pattern of the equine alpha 1(I) chain resembled the guinea pig alpha 1(I) chain, which does not contain methionine between CB7 and CB6.
- The researchers confirmed that purified equine alpha 1(I)CB7,6 contained no methionine, methionine sulfoxide, or homoserine lactone.
- Using mass spectrometry, they determined a mass of 42.26 kd, and established the first 12 amino acids of this CB7,6 were identical to the sequence of human alpha 1(I)CB7 through N-terminal sequence analysis.
Cite This Article
APA
Todhunter RJ, Wootton JA, Lust G, Minor RR.
(1994).
Structure of equine type I and type II collagens.
Am J Vet Res, 55(3), 425-431.
Publication
Researcher Affiliations
- Department of Pathology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.
MeSH Terms
- Amino Acids / analysis
- Animals
- Cartilage, Articular / chemistry
- Chromatography
- Collagen / chemistry
- Collagen / isolation & purification
- Cyanogen Bromide
- Electrophoresis, Polyacrylamide Gel
- Guanidine
- Guanidines
- Horses
- Peptide Fragments / chemistry
- Peptide Fragments / isolation & purification
- Peptide Mapping
- Skin / chemistry
- Tendons / chemistry
Grant Funding
- 07344-01A1 / PHS HHS
- AR 20793 / NIAMS NIH HHS
Citations
This article has been cited 3 times.- Pasculli A, Gurrado A, De Luca GM, Mele A, Marzullo A, Mangone A, Cellamare S, Ferraro V, Maqoud F, Caggiani MC, Rana F, Cavallaro G, Prete FP, Tricarico D, Altomare CD, Testini M. Bridging repair of the abdominal wall in a rat experimental model. Comparison between uncoated and polyethylene oxide-coated equine pericardium meshes. Sci Rep 2020 Apr 24;10(1):6959.
- Sejersen MH, Frost P, Hansen TB, Deutch SR, Svendsen SW. Proteomics perspectives in rotator cuff research: a systematic review of gene expression and protein composition in human tendinopathy. PLoS One 2015;10(4):e0119974.
- Nicoletti A, Fiorini M, Paolillo J, Dolcini L, Sandri M, Pressato D. Effects of different crosslinking conditions on the chemical-physical properties of a novel bio-inspired composite scaffold stabilised with 1,4-butanediol diglycidyl ether (BDDGE). J Mater Sci Mater Med 2013 Jan;24(1):17-35.
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